In this study, we aimed to study the role of miRNAs in intrauterine adhesion (IUA) disease. An IUA cell model was constructed by TGF-β1. Smad3 inhibitor (SIS3) can inhibit the Smad3 signaling pathway and affect the role of TGF-β1; thus, it was used to identify the role of Smad3 and related miRNAs in IUA. Cell number significantly increased in the TGF-β1 group after 72 h and 96 h, respectively, compared with that in the control group (P < 0.05). However, cell proliferation was significantly decreased in the TGF-β1 + SIS3 group (P < 0.0001). Cell apoptosis was increased in the TGF-β1 + SIS3 group compared with that in the TGF-β1 group. Western Blot (WB) analysis suggested that TGF-β1 treatment could effectively increase the expression of α-SMA, COL1, Smad3, and p-Smad3, which could be inhibited by SIS3 treatment. A total of 235 and 530 differentially expressed miRNAs in the TGF-β1 + SIS3 group were significantly up- and downregulated compared with those in the TGF-β1 group, respectively. These differentially expressed miRNAs were enriched in the MAPK and PI3K-AKT pathways. The ten most differentially expressed miRNAs were selected to verify their expressions using quantitative real-time polymerase chain reaction (qPCR). Furthermore, overexpression of rno-miR-3586-3p and rno-miR-455-5p can promote cell proliferation and exacerbate the IUA pathogenic process. However, overexpression of rno-miR-204-3p and rno-miR-3578 can inhibit cell behavior and IUA progression. The above results can provide detailed information for the understanding of IUA molecular mechanisms.

中文翻译：

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miRNA与TGF-β1/ Smad3抑制剂联合在宫内大鼠黏附细胞模型中的功能分析。

在这项研究中，我们旨在研究miRNA在子宫内粘连（IUA）疾病中的作用。用TGF-β1构建IUA细胞模型。Smad3抑制剂（SIS3）可抑制Smad3信号通路并影响TGF-β1的作用。因此，它被用来鉴定IUMA中Smad3和相关miRNA的作用。与对照组相比，TGF-β1组的细胞数在72 h和96 h显着增加（P <0.05）。然而，TGF-β1+ SIS3组的细胞增殖明显减少（P <0.0001）。与TGF-β1组相比，TGF-β1+ SIS3组的细胞凋亡增加。Western Blot（WB）分析表明，TGF-β1处理可有效提高α-SMA，COL1，Smad3和p-Smad3的表达，而SIS3处理可抑制该表达。与TGF-β1组相比，TGF-β1+ SIS3组中共有235和530个差异表达的miRNA分别显着上调和下调。这些差异表达的miRNA富含MAPK和PI3K-AKT途径。使用定量实时聚合酶链反应（qPCR）选择了十个差异最大的miRNA，以验证其表达。此外，rno-miR-3586-3p和rno-miR-455-5p的过表达可以促进细胞增殖并加剧IUA的致病过程。但是，rno-miR-204-3p和rno-miR-3578的过度表达可以抑制细胞行为和IUA进展。以上结果可为了解IUA分子机制提供详细信息。